CN104585027A - Seedling hardening method for ginger tissue culture seedlings - Google Patents
Seedling hardening method for ginger tissue culture seedlings Download PDFInfo
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Abstract
The invention discloses a seedling hardening method for ginger tissue culture seedlings. The method comprises the following steps: (1) selecting healthy and strong fresh ginger without pests as a tissue culture material; disinfecting ginger pieces and burying the disinfected ginger pieces in a sand bed for accelerating germination, sterilizing and stripping 0.3-0.5mm stem tips for inoculating; (2) cultivating the stem tips sequentially by virtue of an aseptic seedling medium, an induction medium, a multiplying medium and a rooting medium so as to obtain tissue culture bottle seedlings capable of leaving a culture room; and (3) uncovering and hardening the seedlings when 4-5 white roots germinates from the tissue culture seedlings in a culture bottle, moving the culture bottle out of the culture room 5 days later, hardening the seedlings under natural illumination at a temperature of 24-26 DEG C for 7 days, taking out the tissue culture seedlings out of the culture bottle after 5-6 leaves which are 5-7cm long appear in the tissue culture seedlings, cleaning the mediums, transplanting the seedlings in a matrix of peat, perlite and vermiculite at the ratio of 2 to 1 to 1, carrying out conventional fertilizer and water management until finished seedlings are obtained, and taking the seedlings out of a nursery. The method disclosed by the invention is simple and easy to implement, low in cost and high in transplanting survival rate.
Description
Technical field
The present invention relates to a kind of breeding method of ginger, particularly relate to a kind of hardening off method of ginger plantlet in vitro.
Background technology
Ginger (Zi ngi ber offici nale Rose) belongs to Zingiber ginger and belongs to herbaceos perennial, is also annual vegetable crop.Ginger is nutritious, and has unique local flavor, can be used as a kind of important flavouring.Ginger also has medical value [1], well received on the market, is thus in great demand.In recent years along with the increase of ginger export trade volume, cultivated area also expands year by year.But ginger carries out vegetative propagation with underground stem tuber, reproduction coefficient low [2-3] for a long time in production.Trophosome plantation for many years causes virus and infection process, and virus [4-6] and the withered Pseudomonas alba of green grass or young crops very easily by kind ginger accumulate in vivo, ginger Different Varieties is caused to be degenerated, vitality declines, and resistance reduces, and easily brings out the multiple diseases such as bacterial wilt of ginger and ginger bark favus, Field diseases can up to 70% ~ 80%, cause product qualitative change bad, pharmaceutical ingredient content is unstable, produces constitute a serious threat to ginger.In recent years, utilize tissue culture technique carry out ginger take off bacterium breeding achieved very ten-strike.In conventional cultivation, get the kind ginger stripping and slicing plantation of rudiment, need plant ginger 4500 ~ 6000kg/hm2, the consumption of planting ginger is very large; Therefore ginger ginger bud tissue is utilized to carry out the mode of tissue cultures in necessarily a kind of trend in future.But, at present about ginger plantlet in vitro cultivation, domestication hardening off method report still relatively little.
Summary of the invention
The object of the present invention is to provide a kind of simple, cost is lower and the hardening off method that ginger plantlet in vitro survival rate is high.
To achieve these goals, present invention employs following technical scheme:
A hardening off method for ginger plantlet in vitro, comprises the following steps successively:
(1) process of seedling tissue:
Select healthy and strong, be tissue cultures material without the ginger of damage by disease and insect, by ginger block with after the 500 times of immersion bubble half an hour sterilizations of 50% carbendazim, be embedded in clean husky bed, carry out vernalization at ambient temperature, as the long 1 ~ 2cm of bud, distilled water flushing is used to remove sandy soil, spire is peelled off at aseptic indoor dissecting needle, get 3 ~ 5mm ginger bud, with 70% alcohol sterilizing one minute, then use 1% ~ 10NaClO sterilizing 20 minutes, distilled water flushing 3 times, superclean bench carries out stem apex stripping with anatomical lens, and the stem apex peelling off 0.3 ~ 0.5mm is inoculated;
(2) cultivation of plantlet in vitro:
The stem apex of 0.3 ~ 0.5mm in step (1) is inoculated in Aseptic seedling culture base, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate, until become explant aseptic seedling when stem apex grows to 1 ~ 2cm; Described Aseptic seedling culture base is: MS+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
Explant aseptic seedling is inoculated in inducing culture, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate and just grow to 1.0 ~ 3.0cm for indefinite bud to be formed in 22 ~ 25 days; Described inducing culture is: MS+ zeatin 2mg/L+6_BA 1.5mg/L+NAA 0.15mg/L+IAA 0.1mg/L+ sucrose 40g/L+ agar 7g/L, pH value is 6.0
Just be inoculated in proliferated culture medium for adventitious shoot by above-mentioned, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate 10 ~ 15 days to forming subculture adventitious shoot and growing to 3.0 ~ 6.0cm; Described proliferated culture medium is: MS+ zeatin 1mg/L+6_BA 2mg/L+NAA 0.25mg/L+IAA 0.3mg/L+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
Above-mentioned subculture adventitious shoot is inoculated in root media, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate 12 ~ 15 days, when seedling grows to 4 ~ 8cm and at least has the root of 3 long >=2cm, can go out the tissue-culture container seedling of culturing room; Described root media is: MS+ gibberellin 1mg/L+KT 2mg/L+NAA 0.1mg/L+IAA 0.05mg/L+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
(3) domestication of plantlet in vitro:
When plantlet in vitro in blake bottle is grown and had 4 ~ 5 white roots, take off lid hardening, and in blake bottle, add a small amount of distilled water; After 5 days, blake bottle is shifted out culturing room, at the condition lower refining seedling 7 days of natural lighting and temperature 24 ~ 26 DEG C, treat that 5 ~ 6 leaves appear in plantlet in vitro, during leaf length 5 ~ 7cm, in blake bottle, take out plantlet in vitro and clean medium, be transplanted into the peat composed of rotten mosses: perlite: in the matrix of vermiculite=2:1:1, rich water quality management is to growing up to finished product seedling routinely, goes out garden.
The present invention have employed the new formula medium being different from conventional medium in the tissue cultures stage; Multiple somatotropin acts synergistically mutually, allows the multiple bud of Ginger Tissue, how long root, and after transplanting, its survival rate height more in the past.
MS medium:
6_BA: i.e. 6-benzyl aminoadenine, is a kind of basic element of cell division, mainly causes cell division, the formation of induced bud and the growth of promotion bud.
NAA: methyl α-naphthyl acetate (1-Naphthaleneacetic acid), be called for short NAA, being a kind of organic compound (chemical formula: C10H7CH2CO2H), is a kind of colorless solid being soluble in organic solvent.Its structure is that No. 1 position of naphthalene replaces with carboxymethyl.It is a kind of plant hormone auxin, is usually used in commercial root of hair powder or rooting agent, uses during cuttage breeding use plant.It also can be used for Plant Tissue Breeding.
IAA: indole-3-acetic acid is ubiquitous Endogenous auxin in a kind of plant corpus, belongs to Benzazole compounds.On a cellular level, growth hormone can stimulate cambial cell to divide; Stimulate the cell elongation of branch, suppress root cell growth; Promote the differentiation of xylem, phloem cell, promote cutting root of hair, regulate the morphogenesis of callus.
KT: chemical name is 6-glycosyl aminopurine (or N6-furfuryladenine), molecular formula C10H9N5O; It is a kind of non-natural basic element of cell division, can promote Cell Differentiation, division, growth; Evoked callus sprouts; Remove apical dominance; Promote seed sprouting, break the dormancy of lateral bud.
Zeatin: a kind of natural basic element of cell division, it extracts from the seed of sweet corn pustulation period and the 1st n cell mitogen going out of crystallization; Promote that callus germinates; Can Prof. Du Yucang.
Gibberellin is the class plant hormone extensively existed.Its chemical constitution belongs to Diterpenes acid, is derived and obtain by Fourth Ring skeleton.It is a new generation's novel high-tech conditioning agent product successfully developed in conjunction with international technology, gibberellin is the improved medicine of Activol, English name: gibberellin, be called for short GA4+7, its effect is absolutely to the efficient of crop, effect is lasting, more efficient, more stable, safer, Seedling Stage starts to spray into the best, well developed root system can be made, pre-disease prevention again, it can promote axis significantly, leaf grows, as vegetative period sprays, also can make balanced in nutrition, contribute to crop growing state, Spraying in the flowering stage, can flower and fruit protecting, also Fruit can be made, more there is U.S. fruit effect, cotton full-bloom stage sprays effectively can reduce fruit abscission rate, improve bolling rate, and efficient solution can remove crop pest
Advantage of the present invention:
1. hardening off method of the present invention is simple, and cost is lower and transplanting survival rate is high.
2. select ginger stem apex as tissue cultures material, ginger plantlet in vitro has that growth is fast, growing way is prosperous, disease-resistant, strong stress resistance, ginger block color and luster cadmium yellow, evenly neat, the characteristic such as pungent is dense, quality better, output are high.
3. in the tissue cultures stage, have employed the synergistic new formula medium of multiple hormone, and be select hormon according to the different stages, compare conventional formulation medium, have better effect.As added zeatin at inducing culture, the multiple bud of Ginger Tissue can be promoted; Add gibberellin at root media, plantlet in vitro the most how long root can be promoted, contribute to improving the survival rate that plantlet in vitro is transplanted.
Embodiment
Below in conjunction with embodiment, the present invention is further described.
Embodiment 1:
A hardening off method for ginger plantlet in vitro, comprises the following steps successively:
(1) process of seedling tissue:
Select healthy and strong, be tissue cultures material without the ginger of damage by disease and insect, by ginger block with after the 500 times of immersion bubble half an hour sterilizations of 50% carbendazim, be embedded in clean husky bed, carry out vernalization at ambient temperature, when bud is about 1cm, distilled water flushing is used to remove sandy soil, spire is peelled off at aseptic indoor dissecting needle, get about 3mm ginger bud, with 70% alcohol sterilizing one minute, then use 1%NaClO sterilizing 20 minutes, distilled water flushing 3 times, superclean bench carries out stem apex stripping with anatomical lens, and the stem apex stripping about 0.3mm is inoculated;
(2) cultivation of plantlet in vitro:
The stem apex of the about 0.3mm in step (1) is inoculated in Aseptic seedling culture base, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate, until become explant aseptic seedling when stem apex grows to about 1cm; Described Aseptic seedling culture base is: MS+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
Explant aseptic seedling is inoculated in inducing culture, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate and just grow to about 1cm for indefinite bud to be formed in 22 days; Described inducing culture is: MS+ zeatin 2mg/L+6_BA 1.5mg/L+NAA 0.15mg/L+IAA 0.1mg/L+ sucrose 40g/L+ agar 7g/L, pH value is 6.0
Just be inoculated in proliferated culture medium for adventitious shoot by above-mentioned, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate about 10 days to forming subculture adventitious shoot and growing to about 3cm; Described proliferated culture medium is: MS+ zeatin 1mg/L+6_BA 2mg/L+NAA 0.25mg/L+IAA 0.3mg/L+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
Above-mentioned subculture adventitious shoot is inoculated in root media, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate 12 days, when seedling grows to about 4cm and at least has the root of 3 long >=2cm, can go out the tissue-culture container seedling of culturing room; Described root media is: MS+ gibberellin 1mg/L+KT 2mg/L+NAA 0.1mg/L+IAA 0.05mg/L+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
(3) domestication of plantlet in vitro:
When plantlet in vitro in blake bottle is grown and had 4 white roots, take off lid hardening, and in blake bottle, add a small amount of distilled water; After 5 days, blake bottle is shifted out culturing room, at the condition lower refining seedling 7 days of natural lighting and temperature 24 DEG C, treat that 5 leaves appear in plantlet in vitro, when leaf is about 5cm, in blake bottle, take out plantlet in vitro and clean medium, be transplanted into the peat composed of rotten mosses: perlite: in the matrix of vermiculite=2:1:1, rich water quality management is to growing up to finished product seedling routinely, goes out garden.
Embodiment 2:
A hardening off method for ginger plantlet in vitro, comprises the following steps successively:
(1) process of seedling tissue:
Select healthy and strong, be tissue cultures material without the ginger of damage by disease and insect, by ginger block with after the 500 times of immersion bubble half an hour sterilizations of 50% carbendazim, be embedded in clean husky bed, carry out vernalization at ambient temperature, when bud is about 2cm, distilled water flushing is used to remove sandy soil, spire is peelled off at aseptic indoor dissecting needle, get 3 ~ 5mm ginger bud, with 70% alcohol sterilizing one minute, then use 10%NaClO sterilizing 20 minutes, distilled water flushing 3 times, superclean bench carries out stem apex stripping with anatomical lens, and the stem apex stripping about 0.5mm is inoculated;
(2) cultivation of plantlet in vitro:
The stem apex of the about 0.5mm in step (1) is inoculated in Aseptic seedling culture base, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate, until become explant aseptic seedling when stem apex grows to about 2cm; Described Aseptic seedling culture base is: MS+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
Explant aseptic seedling is inoculated in inducing culture, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate and just grow to about 3.0cm for indefinite bud to be formed in 25 days; Described inducing culture is: MS+ zeatin 2mg/L+6_BA 1.5mg/L+NAA 0.15mg/L+IAA 0.1mg/L+ sucrose 40g/L+ agar 7g/L, pH value is 6.0
Just be inoculated in proliferated culture medium for adventitious shoot by above-mentioned, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate 15 days to forming subculture adventitious shoot and growing to about 6.0cm; Described proliferated culture medium is: MS+ zeatin 1mg/L+6_BA 2mg/L+NAA 0.25mg/L+IAA 0.3mg/L+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
Above-mentioned subculture adventitious shoot is inoculated in root media, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate 15 days, when seedling grows to about 8cm and at least has the root of 3 long >=2cm, can go out the tissue-culture container seedling of culturing room; Described root media is: MS+ gibberellin 1mg/L+KT 2mg/L+NAA 0.1mg/L+IAA 0.05mg/L+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
(3) domestication of plantlet in vitro:
When plantlet in vitro in blake bottle is grown and had 5 white roots, take off lid hardening, and in blake bottle, add a small amount of distilled water; After 5 days, blake bottle is shifted out culturing room, at the condition lower refining seedling 7 days of natural lighting and temperature 26 DEG C, treat that 6 leaves appear in plantlet in vitro, when leaf is about 7cm, in blake bottle, take out plantlet in vitro and clean medium, be transplanted into the peat composed of rotten mosses: perlite: in the matrix of vermiculite=2:1:1, rich water quality management is to growing up to finished product seedling routinely, goes out garden.
Embodiment 3:
A hardening off method for ginger plantlet in vitro, comprises the following steps successively:
(1) process of seedling tissue:
Select healthy and strong, be tissue cultures material without the ginger of damage by disease and insect, by ginger block with after the 500 times of immersion bubble half an hour sterilizations of 50% carbendazim, be embedded in clean husky bed, carry out vernalization at ambient temperature, as the long 1.5cm of bud, distilled water flushing is used to remove sandy soil, spire is peelled off at aseptic indoor dissecting needle, get 4mm ginger bud, with 70% alcohol sterilizing one minute, then use 5%NaClO sterilizing 20 minutes, distilled water flushing 3 times, superclean bench carries out stem apex stripping with anatomical lens, and the stem apex stripping 0.4mm is inoculated;
(2) cultivation of plantlet in vitro:
The stem apex of the 0.4mm in step (1) is inoculated in Aseptic seedling culture base, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate, until become explant aseptic seedling when stem apex grows to 1.5cm; Described Aseptic seedling culture base is: MS+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
Explant aseptic seedling is inoculated in inducing culture, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate and just grow to 2cm for indefinite bud to be formed in 24 days; Described inducing culture is: MS+ zeatin 2mg/L+6_BA 1.5mg/L+NAA 0.15mg/L+IAA 0.1mg/L+ sucrose 40g/L+ agar 7g/L, pH value is 6.0
Just be inoculated in proliferated culture medium for adventitious shoot by above-mentioned, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate 13 days to forming subculture adventitious shoot and growing to 5cm; Described proliferated culture medium is: MS+ zeatin 1mg/L+6_BA 2mg/L+NAA 0.25mg/L+IAA 0.3mg/L+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
Above-mentioned subculture adventitious shoot is inoculated in root media, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate 13 days, when seedling grows to 6cm and at least has the root of 3 long >=2cm, can go out the tissue-culture container seedling of culturing room; Described root media is: MS+ gibberellin 1mg/L+KT 2mg/L+NAA 0.1mg/L+IAA 0.05mg/L+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
(3) domestication of plantlet in vitro:
When plantlet in vitro in blake bottle is grown and had 5 white roots, take off lid hardening, and in blake bottle, add a small amount of distilled water; After 5 days, blake bottle is shifted out culturing room, at the condition lower refining seedling 7 days of natural lighting and temperature 25 DEG C, treat that 6 leaves appear in plantlet in vitro, during the long 6cm of leaf, in blake bottle, take out plantlet in vitro and clean medium, be transplanted into the peat composed of rotten mosses: perlite: in the matrix of vermiculite=2:1:1, rich water quality management is to growing up to finished product seedling routinely, goes out garden.
Claims (1)
1. a hardening off method for ginger plantlet in vitro, is characterized in that, comprises the following steps successively:
(1) process of seedling tissue:
Select healthy and strong, be tissue cultures material without the ginger of damage by disease and insect, by ginger block with after the 500 times of immersion bubble half an hour sterilizations of 50% carbendazim, be embedded in clean husky bed, carry out vernalization at ambient temperature, as the long 1 ~ 2cm of bud, distilled water flushing is used to remove sandy soil, spire is peelled off at aseptic indoor dissecting needle, get 3 ~ 5mm ginger bud, with 70% alcohol sterilizing one minute, then use 1% ~ 10NaClO sterilizing 20 minutes, distilled water flushing 3 times, superclean bench carries out stem apex stripping with anatomical lens, and the stem apex stripping 0.3 ~ 0.5mm is inoculated;
(2) cultivation of plantlet in vitro:
The stem apex of 0.3 ~ 0.5mm in step (1) is inoculated in Aseptic seedling culture base, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate, until become explant aseptic seedling when stem apex grows to 1 ~ 2cm; Described Aseptic seedling culture base is: MS+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
Explant aseptic seedling is inoculated in inducing culture, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate and just grow to 1.0 ~ 3.0cm for indefinite bud to be formed in 22 ~ 25 days; Described inducing culture is: MS+ zeatin 2mg/L+6_BA 1.5mg/L+NAA 0.15mg/L+IAA 0.1mg/L+ sucrose 40g/L+ agar 7g/L, pH value is 6.0
Just be inoculated in proliferated culture medium for adventitious shoot by above-mentioned, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate 10 ~ 15 days to forming subculture adventitious shoot and growing to 3.0 ~ 6.0cm; Described proliferated culture medium is: MS+ zeatin 1mg/L+6_BA 2mg/L+NAA 0.25mg/L+IAA 0.3mg/L+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
Above-mentioned subculture adventitious shoot is inoculated in root media, 25 DEG C, light intensity 1500Lx, illumination 12h/d condition of tissue culture under cultivate 12 ~ 15 days, when seedling grows to 4 ~ 8cm and at least has the root of 3 long >=2cm, can go out the tissue-culture container seedling of culturing room; Described root media is: MS+ gibberellin 1mg/L+KT 2mg/L+NAA 0.1mg/L+IAA 0.05mg/L+ sucrose 40g/L+ agar 7g/L, and pH value is 6.0;
(3) domestication of plantlet in vitro:
When plantlet in vitro in blake bottle is grown and had 4 ~ 5 white roots, take off lid hardening, and in blake bottle, add a small amount of distilled water; After 5 days, blake bottle is shifted out culturing room, at the condition lower refining seedling 7 days of natural lighting and temperature 24 ~ 26 DEG C, treat that 5 ~ 6 leaves appear in plantlet in vitro, during leaf length 5 ~ 7cm, in blake bottle, take out plantlet in vitro and clean medium, be transplanted into the peat composed of rotten mosses: perlite: in the matrix of vermiculite=2:1:1, rich water quality management is to growing up to finished product seedling routinely, goes out garden.
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| CN107278894A (en) * | 2017-07-11 | 2017-10-24 | 永州职业技术学院 | Brave pawl ginger tissue-culturing quick-propagation culture medium |
| CN107810857A (en) * | 2017-12-22 | 2018-03-20 | 济南亚龙生物科技有限公司 | A kind of detoxicated ginger method for tissue culture |
| CN108849355A (en) * | 2018-07-24 | 2018-11-23 | 天峨县平昌生态农业有限公司 | A kind of ginger cultivation method |
| CN111771725A (en) * | 2020-07-31 | 2020-10-16 | 潍坊兴旺生物种业有限公司 | Optimized ginger virus-free seedling regeneration propagation method and development of industrialization process thereof |
| CN115769786A (en) * | 2022-11-30 | 2023-03-10 | 安徽农业大学 | Method for obtaining regeneration seedlings of drynaria fortunei by alternative tissue culture |
| CN115777531A (en) * | 2022-09-19 | 2023-03-14 | 一禾亿株生物种业(深圳)有限公司 | Ginger tissue culture seedling method |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107278894A (en) * | 2017-07-11 | 2017-10-24 | 永州职业技术学院 | Brave pawl ginger tissue-culturing quick-propagation culture medium |
| CN107810857A (en) * | 2017-12-22 | 2018-03-20 | 济南亚龙生物科技有限公司 | A kind of detoxicated ginger method for tissue culture |
| CN108849355A (en) * | 2018-07-24 | 2018-11-23 | 天峨县平昌生态农业有限公司 | A kind of ginger cultivation method |
| CN111771725A (en) * | 2020-07-31 | 2020-10-16 | 潍坊兴旺生物种业有限公司 | Optimized ginger virus-free seedling regeneration propagation method and development of industrialization process thereof |
| CN115777531A (en) * | 2022-09-19 | 2023-03-14 | 一禾亿株生物种业(深圳)有限公司 | Ginger tissue culture seedling method |
| CN115769786A (en) * | 2022-11-30 | 2023-03-10 | 安徽农业大学 | Method for obtaining regeneration seedlings of drynaria fortunei by alternative tissue culture |
| CN115769786B (en) * | 2022-11-30 | 2023-12-19 | 安徽农业大学 | Method for obtaining copper tomb-white Jiang Zaisheng seedlings in alternating tissue culture mode |
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